期刊文献+
共找到2篇文章
< 1 >
每页显示 20 50 100
牛肠激酶基因工程菌的构建、高密度发酵、纯化及活性鉴定 被引量:5
1
作者 李德华 苟兴华 +4 位作者 胡海洋 徐琦 刘忠荣 吴洽庆 余晓东 《应用与环境生物学报》 CAS CSCD 北大核心 2006年第6期819-823,共5页
肠激酶(enterokinase,EK)是一种专一识别DDDDK氨基酸序列、并水解K后肽键的丝氨酸蛋白水解酶.根据GenBank序列进行人工合成牛肠激酶轻链基因cDNA,测序正确后将其插入甲醇酵母分泌型载体pPICZαA,得到重组质粒pPICZαA/EKL,重组载体经线... 肠激酶(enterokinase,EK)是一种专一识别DDDDK氨基酸序列、并水解K后肽键的丝氨酸蛋白水解酶.根据GenBank序列进行人工合成牛肠激酶轻链基因cDNA,测序正确后将其插入甲醇酵母分泌型载体pPICZαA,得到重组质粒pPICZαA/EKL,重组载体经线性化后转化Pichia pastorisSMD1168H,筛选得到重组牛肠激酶轻链工程菌.分别对重组酵母工程菌进行高密度发酵、rEKL(recombinant enterokinase light chain,rEKL)纯化、N端序列测定、生物学活性鉴定等研究工作.结果表明,发酵上清中rEKL含量高,纯化的rEKL专一性强、无其他杂酶活性、N端15个氨基酸序列与文献报道一致. 展开更多
关键词 肠激酶(enterokinase EK) 甲醇酵母(Pichia pastoris) 高密度发酵
下载PDF
Western blotting in the diagnosis of duodenal-biliary and pancreaticobiliary refluxes in biliary diseases 被引量:6
2
作者 Chun-Chih Chen 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2009年第6期608-613,共6页
BACKGROUND:Currently adopted diagnostic methods for duodenal-biliary and pancreaticobiliary refluxes carry many flaws,so the incidence of the two refluxes demands further larger sample size studies.This study aimed to... BACKGROUND:Currently adopted diagnostic methods for duodenal-biliary and pancreaticobiliary refluxes carry many flaws,so the incidence of the two refluxes demands further larger sample size studies.This study aimed to evaluate Western blotting for the diagnosis of refluxes in biliary diseases.METHODS:An oral radionuclide 99m Tc-DTPA test(radio-nuclide,RN)was conducted for the observation of duodenal-biliary reflux prior to measuring bile radioactivity and Western blotting for detecting bile enterokinase(EK).Pancreaticobiliary reflux was assessed by biochemical and Western blotting tests for biliary amylase activity and trypsin-1,respectively.In accordance with bile sample origin,our samples were classified into ductal bile and gall bile groups;based on each individual biliary disease,we further classified the ductal bile group into five subgroups,and the gall bile group into four sub-groups.Western blotting was conducted to assess the two refluxes in biliary diseases.RESULTS:Consistencies were noted between EK and RN tests when diagnosing duodenal-biliary reflux(P<0.001).The amylase and trypsin-1 tests also showed consistency in diagnosing pancreaticobiliary reflux(P<0.001).Amylase and lipase levels within gall and ductal bile were strongly correlated(P<0.05).In the common bile duct pigment stone group,the EK and trypsin-1 positive rates were found to be insignificant(P>0.05);in the common bile duct cyst group,the EK positive rate was significantly lower than the trypsin-1 positive rate(P<0.05).CONCLUSIONS:Western blotting can accurately reflect duodenal-biliary and pancreaticobiliary refluxes.EK has greater sensitivity than RN for duodenal-biliary reflux.The majority of biliary amylase and lipase comes from the pancreas in all biliary diseases;pancreaticobiliary reflux is the predominant source in the common bile duct cyst group and duodenal-biliary reflux is responsible for the ductal pigment stone group. 展开更多
关键词 pancreaticobiliary REFLUX duodenal-biliary REFLUX trypsin-1 enterokinase biliary diseases
下载PDF
上一页 1 下一页 到第
使用帮助 返回顶部